Unmet Need
Gene therapy has become increasingly popular for treating genetic and acquired diseases, with nearly 1000 ongoing clinical trials globally. These therapies are typically delivered via viral vectors, which are produced in vector production cell lines following transient transfection using nanoparticles such as those made of plasmid DNAs (pDNA) and polyethyleneimine (PEI), a polycation. With an increasing number of clinical trials and studies, the production demand of viral vectors has increased substantially. However, the current standard processes for preparing cell transfection nanoparticles require batch-mode production immediately before cell transfection for vector manufacturing. This pDNA/polycation nanoparticle preparation process is prone to lower consistency at the industry manufacturing scale, resulting in higher variability in production yield of viral vectors, comparing to the bench tests. There is a need for a low-variability, high transfection efficiency, and off-the-shelf pDNA/polycation nanoparticles that can enable large scale manufacturing of viral vectors.
Technology Overview
Johns Hopkins researchers have developed a scalable method of producing shelf-stable pDNA/polycation particles in the range of 50 to 1000 nanometers. They have also determined the optimal composition and size of the pDNA/polycation particles for maximum in vitro transfection efficiency, with an optimal size of 400 to 500 nanometers. The off-the-shelf formulation of the 400-nm pDNA/PEI particles demonstrated superior shelf stability at -80 degrees Celsius with preserved physical properties and transfection activity. The particle formulation was validated using lentiviral vectors and demonstrated consistent yield equivalent to standard manual preparation methods, drastically streamlining the viral manufacturing process and enabling reliable production of viral vectors. In addition, these shelf-stable, off-the-shelf DNA/polycation particles yield superior and reproducible transfection activity. Beyond viral vector production, these nanoparticles can be used for ex vivo transfection of cells for regenerative therapy and immunotherapy.
Stage of Development
Method development and validation completed.
Patent
Provisional patent filed.
